Browsing by Subject "Interleucina-1"

Now showing 1 - 3 of 3
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    Interleukin-1β isolated from a marine fish reveals up-regulated expression in macrophages following activation with lipopolysaccharide and lymphokines
    (Elsevier, 2001) Pelegrin, Pablo; García-Castillo, Jesús; Meseguer, José; Mulero Méndez, Victoriano Francisco; Bioquímica y Biología Molecular B e Inmunología
    The gilthead seabream IL-1β gene consists of five exons/four introns. The complete coding sequence contains a 102 bp 5’ untraslated region (UTR), a single open reading frame of a 762 bp which translates into a 253 amino acid molecule, and a 407 bp 3’UTR with a polyadenilation signal 14 nucleotides upstream of the poly(A)tail. The seabream sequence has highest degree of nucleotide (61.7%) and amino acid (53%) identity with the trout IL-1β sequences. The IL-1β message was detected by RT-PCR in head-kidney, blood, spleen, liver, gill and peritoneal exudate of both non-infected and Vibrio anguillarum-challenged fish. More importantly, IL-1β was highly expressed by purified macrophage monolayers and was up-regulated by lipopolysaccharide and lymphocyte-derived macrophage-activating factor stimulation.
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    Phylogeny of cytokines: molecular cloning and expression analysis of sea bass Dicentrarchus labrax interleukin-1b
    (2001) Scapigliati, G.; Buonocore, F.; Bird, S.; Zou, J.; Falasca, C.; Prugnoli, D.; Secombes, C. J.; Pelegrín Vivancos, Pablo; Bioquímica y Biología Molecular B e Inmunología
    In this paper the cloning of interleukin-1b (IL-1b) from the fish Dicentrarchus labrax (sea bass) is described. Using degenerate primers designed from known IL-1b sequences, a cDNA fragment was amplified by PCR and elongated by 3’ and 5’ RACE to give the full-length coding sequence for sea bass IL-1. The cDNA is 1292 bp, lacks a putative ICE cut site, and codes for a deduced peptide of 29.4 kDa with a pI of 5.1. Sequence analysis showed highest amino acid similarity with rainbow trout (62%), Xenopus (46%), and carp (45.5%) IL-1b sequences. Expression studies show that sea bass IL-1b can be upregulated by bacterial lipopolysaccharide both in vitro and in vivo in leucocytes from blood, head-kidney, spleen, gills and liver, whereas the IL-1b transcript was not detectable in thymus and gut-associated lymphoid tissue. Northern blot analysis with head-kidney leucocyte RNA showed a main LPS- upregulated band at 1.3 kb, and two minor bands at 0.9 and 3.0 kb, respectively. Phylogenetic comparisons with IL-1b from other vertebrates is presented.
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    Production and mechanism of secretion of interleukin-1b from the marine fish gilthead seabream
    (2004) Elena, Chaves-Pozo; José, Meseguer; Mulero Méndez, Victoriano Francisco; Pelegrín Vivancos, Pablo; Bioquímica y Biología Molecular B e Inmunología
    Interleukin-1b (IL-1b) is a secretory cytokine lacking a signal peptide, and does not follow the classical endoplasmic reticulum to Golgi pathway of secretion. Its post-translational processing by IL-1b-converting enzyme (ICE) and subsequent release from activated macrophages requires ATP acting on P2X7 receptors. No information is available on the production and release of fish IL-1b, but the IL-1b gene sequences reported to date lack a conserved ICE recognition site. We show for the first time that lipopolysaccharide (LPS)/macrophage-activating factor (MAF)/bacterial DNA (VaDNA)-primed immune cells of fish accumulate intracellular IL-1b as a ~30 kDa polypeptide (proIL-1b). The combination of LPS and VaDNA was found to be synergistic, suggesting that each ligand is recognized by a different pattern recognition receptor (PRR). More importantly, addition of extracellular ATP does not promote IL-1b secretion by immune cells and fails to induce phosphatidylserine (PS) flip. In contrast, fish SAF-1 fibroblasts shed microvesicles containing a 22 kDa IL-1b form within 30 min of activation with ATP. Notably, the post-translational processing of IL-1b by SAF-1 cells is abrogated by a specific ICE inhibitor.